LSD1 collaborates with EZH2 to regulate expression of interferon-stimulated genes
LSD1 collaborates with EZH2 to regulate expression of interferon-stimulated genes
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LSD1 与 EZH2 合作调节干扰素刺激基因的表达。
DOI：
10.1016/j.biopha.2017.01.055
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发表时间：
2017-04-01
影响因子：
7.5
通讯作者：
Hu, Xin
中科院分区：
文献类型：
--
作者：
Jin, Yue;Huo, Bo;Hu, Xin
组蛋白甲基化是一种复杂的、动态的表观遗传修饰，它调控着基因转录、染色体结构和细胞分化。在这里，我们发现了H3 K4脱甲基酶，赖氨酸特异性脱甲基酶1（LSD 1，CoREST阻遏物复合物的重要组成部分）和H3 K27甲基转移酶，zeste同源物2的增强子（EZH 2，PRC 2的必要组成部分）之间的相互作用。采用免疫沉淀法和GST-pull down法观察蛋白质间的相互作用。培养MCF-7细胞并用siRNA转染。采用实时荧光定量聚合酶链反应（RT-PCR）和蛋白质免疫印迹法（western blot）分别检测mRNA和蛋白质的表达。进行HPLC和LC-MS/MS分析以纯化蛋白质。进行基于RTPCR的ChIP定量分析。MCF-7细胞中LSD 1与组蛋白修饰蛋白EZH 2相互作用。LSD 1和EZH 2靶向一些共同的基因。LSD 1敲低和EZH 2敲低影响蛋白质表达。LSD 1敲低和EZH 2敲低影响参与IFN信号通路的蛋白质。LSD 1和EZH 2修饰IRF 9基因位点的组蛋白甲基化。我们用蛋白质组学方法系统地分析了受LSD 1或EZH 2敲低影响的蛋白质，并确定了干扰素途径和一些其他途径通常受到影响。LSD 1和EZH 2之间的相互作用稳定了LSD 1与IRF 9启动子区的结合，IRF 9是干扰素途径的关键转录因子。因此，我们的研究表明，组蛋白去甲基化酶和甲基转移酶之间的协调可能作为一个双锁系统，以抑制干扰素刺激基因的表达。(C)2017年由Elsevier Masson SAS出版。
Histone methylation is a complicate and dynamic epigenetic modification that regulates gene transcription, chromosomal structure and cell differentiation. Here, we discovered the interaction between the H3K4 demethylase, lysine specific demethylase 1 (LSD1, an important component of CoREST repressor complex) and the H3K27 methyltransferase, enhancer of zeste homolog 2 (EZH2, an essential component of PRC2). Immuno-precipitation and GST-pull down assay were performed to observe the interaction between the proteins. The MCF-7 cells were cultured and transfected with the siRNA. The mRNA and proteins were examined by using the real-time polymerase chain reaction (RT-PCR) and western blot assay, respectively. HPLC and LC-MS/MS analysis were performed to purify the proteins. RTPCR- based quantitative ChIP analysis were performed. LSD1 interacts with histone modification protein EZH2 in MCF-7 cells. LSD1 and EZH2 target a few common genes. LSD1 knockdown and EZH2 knockdown affect protein expression. LSD1 knockdown and EZH2 knockdown affect the proteins involving in IFN signaling pathway. LSD1 and EZH2 modify histone methylation at IRF9 gene locus. We systematically analyzed the proteins that are affected by either LSD1 or EZH2 knockdown with proteomic approaches and identified that the interferon pathway and some other pathways are commonly affected. The interaction between LSD1 and EZH2 stabilizes the binding of LSD1 to the promoter region of IRF9, which is a key transcription factor of the interferon pathway. In conclusion, our study revealed that the coordination between histone demethylases and methyl-transferases might serve as a double lock system to suppress the expression of interferon stimulated genes. (C) 2017 Published by Elsevier Masson SAS.